Aging is associated with declines in a number of cognitive domains, particularly memory. As people over the age of 65 will comprise more than 20% of the US population by the year 2040, there is a pressing need to elucidate the mechanisms by which aging impairs memory and to identify improved routes of therapeutic intervention. Importantly, neuropsychological assessments of humans demonstrate that not all aged individuals will be clinically diagnosed with memory impairment, demonstrating that chronological and cognitive aging are not synonymous processes. Unlike Alzheimer's disease, age-related changes in memory are not associated with wide-spread neuronal loss;rather there is a selective decrease in the numbers of synapses within the hippocampus and decreased responsivity of post-synaptic receptors. These behavioral and neuronal sequelae apparent in aged humans can be successfully modeled in aged rats tested for memory of spatial locations. This project will make use of a rodent model of aging integrating aspects of chronological and cognitive aging to investigate associated changes in presynaptic and postsynaptic substrates. Parallel analysis of glutamatergic, cholinergic and GABAergic hippocampal circuits and receptors will determine if changes are specific to a particular neurotransmitter system or more general changes in the aged brain. Changes to innervation of the hippocampus will be visualized by immunohistochemical staining techniques and quantified using unbiased stereological approaches. M1 muscarinic- and mGluR5 metabotropic glutmate receptor-mediated GTP-binding, a functional measure of receptor coupling, will be selectively measured using a scintillation proximity counting technique specific to their cognate G-protein 1-subunit, Gq/11. These changes will be contrasted by measures of GABAB receptor-mediated binding which do not deteriorate with age, but signal via a Gi/Go cascade. In the absence of overt loss of receptor and G-proteins, it has been hypothesized that molecular processes associated with oxidative stress in the aged hippocampus may interfere with normal signal transduction. To test this hypothesis, receptors and G-proteins will be analyzed by immunoprecipitation and mass spectrometry to determine if these specific proteins are oxidatively damaged. Collectively, this project will evaluate age-related changes in three neurotransmitter systems closely tied to normal memory function and determine the mechanisms by which these circuits and synapses are changed in aging. The data derived from these experiments will not only greatly enhance understanding of brain aging and cognition, but elucidate changes to specific neural substrates that are of interest for potential therapeutic modulation by the next generation of pharmaceutical treatments. Furthermore, the diversity of experimental approaches proposed to address these focused research efforts will provide for an excellent pre-doctoral training experience to the applicant in support of his stated career goals and objectives. PUBLIC HEALTH RELEVANCE: As people over the age of 65 will comprise more than 20% of the U.S. population by 2040, appreciating the mechanisms by which memory declines in the elderly is matter of pressing scientific and clinical concern. This research project will measure presynaptic and postsynaptic aspects of neural signaling to determine specific changes to metabotropic receptors and G-proteins in the aged hippocampus associated with memory deficits. The findings of this project will enhance our understanding of the aged brain and provide new insights for therapeutic treatment potential.